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. 2017 Nov 7;114(47):E10112–E10121. doi: 10.1073/pnas.1708367114

Fig. 4.

Fig. 4.

Atg20 is a posttranslationally modified protein. (A) Schematic domain representation of Atg20 with all PTM sites that were experimentally detected by LC-MS/MS analysis. (B) Functionality of Atg20 variants in the Cvt pathway analyzed by the prApe1 maturation assay. The SEY6210 atg20Δ strain was transformed with the plasmids pCuGFP(426), pCuGFP-Atg20(426), pCuGFP-Atg20[10STA](426), and pCuGFP-Atg20[4KR](426). Cells were cultured in rich selective medium. (C) Autophagy induction examined for WT and PTM mutants of Atg20 using the prApe1 maturation assay. The SEY6210 atg20∆ vac8∆ strain was transformed with the same plasmids as indicated in B. Cells were cultured in rich selective medium and then shifted to SD-N medium for 0.5 h. The atg11Δ (SEY6210) strain served as a negative control, and Pgk1 served as a loading control. Quantification of the Ape:prApe1 ratio was determined from three independent experiments. Error bars represent SDs. Statistical significance was tested using the unpaired two-tailed Student’s t test: ***P < 0.005.